Anti-flavivirus compounds and methods of use

ABSTRACT

The present invention concerns the use of compounds and compositions for the treatment or prevention of Flavivirus infections, such as dengue virus infections and Zika virus infections. Aspects of the invention include methods for treating or preventing Flavivirus virus infection, such as dengue virus and Zika virus infection, by administering a compound or composition of the invention, to a subject in need thereof; methods for inhibiting Flavivirus infections, such as dengue virus and Zika virus infections, in a cell in vitro or in vivo; pharmaceutical compositions; packaged dosage formulations; and kits useful for treating or preventing Flavivirus infections, such as dengue virus and Zika virus infections.

CROSS-REFERENCE TO RELATED APPLICATION

The present application claims the benefit of U.S. ProvisionalApplication Ser. No. 62/781,687, filed Dec. 19, 2018, which is herebyincorporated by reference herein in its entirety, including any figures,tables, nucleic acid sequences, amino acid sequences, or drawings.

BACKGROUND OF THE INVENTION

Flaviviruses are a genus within the Flaviviridae family with asingle-stranded, positive-sense RNA genome. Many representatives of thegenus, such as Dengue virus (DENV), West Nile virus (WNV), Zika viruses(ZIKV), tick-borne encephalitis virus, and yellow fever virus, areassociated with pathogenic effects in humans. Currently, approvedtreatments for DENV or ZIKV infection are supportive, but notanti-viral.

ZIKV, a mosquito-borne flavivirus, has re-emerged and spread across theWestern Hemisphere in the past year. First isolated in 1947 from asentinel rhesus macaque in the Ziika Forest region of Uganda [1], ZIKVhad remained in relative obscurity for many years until outbreaks in thePacific islands and then the Americas in the past decade [2-4]. A largeoutbreak started in Brazil in late 2014 and is a growing public healthconcern [5]. Currently, active transmission has been reported in 58countries and territories globally. About 20% of ZIKV infectedindividuals develop symptoms, which mostly resemble symptoms caused byother arboviruses, such as dengue viruses or chikungunya virus. Unlikethese viruses, however, ZIKV causes congenital defects, includingmicrocephaly [6,7], and is also associated with Guillain-Barré syndromein infected adults [8,9].

DENV, the cause of dengue fever, has increased dramatically in recentdecades, becoming one of the most impactful mosquito-borne humanpathogens posing a threat to tropical countries and their visitors.Dengue disease is a global public health threat caused by the spread offour antigenically distinct serotypes of DENV (DENV-1, -2, -3, and -4).Dengue disease can range in severity from mild dengue fever to severe,life-threatening syndromes, dengue hemorrhagic fever (DHF), and dengueshock syndrome (DSS). Infection by one type of DENV can provide immunityagainst that specific serotype, but a subsequent infection by any of theother serotypes of the virus increases the risk of developing severedengue disease, DHF, and DSS. There is currently no specific treatmentfor dengue disease, and most forms of therapy are supportive in nature.

BRIEF SUMMARY OF THE INVENTION

A computational model has been developed based on data from theflavivirus Zika NS-1 assay and was used to predict new anti-Flaviviruscompounds. Because the NS-1 protein is synthesized only in theflavivirus replication stage, the inhibition of NS-1 protein level bycompounds determined in this NS-1 assay, indicates the inhibition ofvirus replication in human cells. The model identified 19 compounds thatwere also experimentally verified to be active and potent in inhibitingNS-1 production and blocking viral replication in human cells. Thesecompounds may be used in new therapies for the treatment of infectionwith flaviviruses, such as Zika virus and dengue virus.

The invention concerns compounds selected from the group consisting ofNCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01 (the chemicalstructures of which are shown in Table 1), or a prodrug, metabolite, orderivative of any of the foregoing, or a pharmaceutically acceptablesalt of any of the foregoing.

Another aspect of the invention is a composition comprising at least onecompound selected from the group consisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; and a pharmaceuticallyacceptable carrier or diluent.

Another aspect of the invention concerns a method for treating orpreventing flavivirus infection in a human or non-human animal subjectin need thereof, the method comprising administering to the subject aneffective amount of at least one compound selected from the groupconsisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; or a composition comprising atleast one of the aforementioned compounds, and pharmaceuticallyacceptable carrier or diluent.

Another aspect of the invention is a method for inhibiting flavivirusinfection in human or non-human animal cells in vitro or in vivo, themethod comprising contacting an effective amount of at least onecompound to a human or non-human animal cell in vitro or in vivo beforeor after exposure of the cell to flavivirus, wherein the compound isselected from the group consisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; or a composition comprising atleast one of the aforementioned compounds, and pharmaceuticallyacceptable carrier or diluent.

Another aspect of the invention concerns a packaged dosage formulationcomprising at least one anti-flavivirus one compound in apharmaceutically acceptable dosage in one or more packages, packets, orcontainers.

Another aspect of the invention concerns a kit comprising, in one ormore containers, at least one anti-flavivirus compound.

DETAILED DESCRIPTION OF THE INVENTION

The flavivirus NS-1 protein is only expressed during the virusreplication stage. The inventors identified several compounds thatinhibited NS-1 production in human cells. These compounds blockflavivirus virus replication and thus may be used as anti-flavivirusagents to inhibit flavivirus infections, such as dengue virus (DENV),West Nile virus (WNV), Zika viruses (ZIKV), tick-borne encephalitisvirus, and yellow fever virus, and others.

An aspect of the invention includes the compounds selected from thegroup consisting of NCGC00102779-01, NCGC00113159-01, NCGC00104879-01,NCGC00025125-18, NCGC00018238-09, NCGC00112058-01, NCGC00169957-03,NCGC00387651-01, NCGC00179895-03, NCGC00108525-01, NCGC00071621-03,NCGC00131231-01, NCGC00246910-02, NCGC00108581-01, NCGC00107055-01,NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01,or a prodrug, metabolite, or derivative of any of the foregoing, or apharmaceutically acceptable salt of any of the foregoing.

The chemical structures of NCGC00102779-01, NCGC00113159-01,NCGC00104879-01, NCGC00025125-18, NCGC00018238-09, NCGC00112058-01,NCGC00169957-03, NCGC00387651-01, NCGC00179895-03, NCGC00108525-01,NCGC00071621-03, NCGC00131231-01, NCGC00246910-02, NCGC00108581-01,NCGC00107055-01, NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, andNCGC00378623-01 are shown in Table 1.

TABLE 1 Sample ID and IUPAC Name Structure 1 NCGC00018238-09 methyl(6-propoxy-1H-benzo[d]imidazol- 2-yl)carbamate

2 NCGC00025125-18 N-[(7S)-1,2,3,10-tetramethoxy-9-oxo-6,7-dihydro-5H-benzo[a]heptalen-7- yl]acetamide

3 NCGC00071621-03 4-(3-(2-methoxyphenyl)-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazol-6- yl)-N,N-dimethylaniline

4 NCGC00072088-02 N-(pyridin-3-yl)thiophene-2-carboxamide

5 NCGC00102779-01 5-(4-ethoxyphenyl)-4-(3-fluorophenyl)-3-(thiophen-2-yl)-4,5- dihydropyrrolo[3,4-c]pyrazol-6(1H)-one

6 NCGC00104879-01 N-(2-chlorobenzyl)-3-(2-methoxyethyl)-2,4-dioxo-1,2,3,4-tetrahydroquinazoline- 7-carboxamide

7 NCGC00107055-01 9-(3-bromo-4,5-dimethoxyphenyl)-2,3,8,9-tetrahydro-[1,4]dioxino[2,3-g]quinolin- 7(6H)-one

8 NCGC00108525-01 ethyl 4-(3-(4-methoxyphenyl)-6-oxo-4-(p-tolyl)-4,6-dihydropyrrolo[3,4-c]pyrazol- 5(1H)-yl)benzoate

9 NCGC00108581-01 ethyl 4-(3-(furan-2-yl)-6-oxo-4-(p-tolyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)- yl)benzoate

10 NCGC00112058-01 N-(2-chlorobenzyl)-1,4,11-trimethyldibcnzo[b,f][1,4]thiazepine-8- carboxamide

11 NCGC00113159-01 methyl 4-[[5-methyl-2-(4-methylphenyl)-1,3-oxazol-4-yl]methyl]furo[3,2-b]pyrrole- 5-carboxylate

12 NCGC00131231-01 4-(tert-butyl)-N-(1-(thiophene-2-carbonyl)-1,2,3,4-tetrahydroquinolin-6-yl)benzamide

13 NCGC00169957-03 3-(sec-butyl)-16-isobutyl-6-isopropyl-5,8,9-trimethyldodecahydropyrrolo[1,2-d][1]oxa[4,7,10,13,16]pentaazacyclo- nonadecine-1,4,7,10,14,17(11H,16H)-hexaone

14 NCGC00179895-03 ((2S,2′R,3′R,4′S,5′S,5a′R,9a′R)-4′-acetoxy-3′-hydroxy-5′,8′-dimethyl-2′,3′,4′,5′,7′,9a′-hexahydrospiro[oxirane-2,10′- [2,5]methanobenzo[b]oxepin]-5a′(6′H)-yl)methyl acetate

15 NCGC00246910-02 7-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one

16 NCGC00263862-02 5-fluoro-1-((4R,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2- yl)pyrimidine-2,4(1H,3H)-dione

17 NCGC00357393-02 (2S,3R,4S,4aR)-2,3,4,7-tetrahydroxy-3,4,4a,5-tetrahydro- [1,3]dioxolo[4,5-j]phenanthridin- 6(2H)-one

18 NCGC00378623-01 (S)-2-((S)-2-(dimethylamino)-3-methylbutanamido)-N-((3R,4S,5S)-3- methoxy-1-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((S)-2-phenyl-1- (thiazol-2-yl)ethyl)amino)propyl)pyrrolidin-1-yl)-5- methyl-1-oxoheptan-4-yl)-N,3-dimethylbutanamide

19 NCGC00387651-01 (2S,2′R,3′R,4′S,5′S,5a′R,7′S,9a′R)-4′-acetoxy-5a′-(acetoxymethyl)-3′-hydroxy-5′,8′-dimethyl-2′,3′,4′,5′,5a′,6′,7′,9a′- octahydrospiro[oxirane-2,10′-[2,5]methanobenzo[b]oxepin]-7′-yl 3- methylbutanoate

Another aspect of the invention includes a composition comprising acompound selected from the group consisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; and a pharmaceuticallyacceptable carrier or diluent. The composition may include one, two ormore compounds of the foregoing compounds. Optionally, the compositionmay include one or more additional biologically active agents, such asan additional agent useful for the treatment or prevention of aflavivirus infection.

Another aspect of the invention is a method for treating or preventingflavivirus infection in a human or non-human animal subject in needthereof, the method comprising administering to the subject an effectiveamount of at least one compound selected from the group consisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; or a composition comprising atleast one of the aforementioned compounds, and pharmaceuticallyacceptable carrier or diluent. In some embodiments, two or more of theaforementioned compounds are administered, within the same formulation,or separate formulations.

In some embodiments, the flavivirus infection is a Zika virus infection.In some embodiments, the flavivirus infection is a dengue virusinfection. The DENV may be any type. For example, the DENV may be any ofserotypes DENV-1, -2, -3, or -4, which follow the human cycle, orDENV-5, which follows the sylvatic cycle (Mustafa Lt Col MS et al.,Medical Journal Armed Forces India, 2015, 71:67-70).

The compounds and compositions may be administered as a therapy, whereinthe subject has the flavivirus infection at the time the compound orcomposition is administered, or the compound or composition may beadministered as prophylaxis, to prevent or delay onset of the flavivirusinfection, wherein the subject does not have the flavivirus infection atthe time the compound or composition is administered.

Optionally, for therapy, the method may include the step of identifyingthe subject as having the flavivirus infection, before administering thecompound or composition. Subjects with a flavivirus infection may beidentified by methods known in the art, such as by assaying a biologicalsample (e.g., blood, serum, or plasma) obtained from the subject for thepresence of flavivirus nucleic acids or flavivirus proteins. Such assaysmay involve, for example, the use of reverse transcriptase-polymerasechain reaction (RT-PCR), immunological assay, or Plaque-reductionneutralization testing (PRNT).

The compounds and compositions may be administered by any effectivemethod. In some embodiments, the compounds and compositions areadministered orally, nasally, rectally, parenterally, subcutaneously,intramuscularly, or intravascularly.

Optionally, the method may further include administering one or moreadditional biologically active agents, such as additional agents fortreating or preventing flavivirus infection, or a symptom thereof, inthe same formulation as the aforementioned anti-flavivirus compound, orin a separate formulation before, during, or after administration of theanti-flavivirus compound.

Another aspect of the invention is a method for inhibiting flavivirusinfection in human or non-human animal cells in vitro or in vivo, themethod comprising contacting an effective amount of at least onecompound to a human or non-human animal cell in vitro or in vivo beforeor after exposure of the cell to flavivirus, wherein the compound isselected from the group consisting of:

NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing; or a composition comprising atleast one of the aforementioned compounds, and pharmaceuticallyacceptable carrier or diluent. In some embodiments, the cell iscontacted with two or more of the aforementioned compounds, within thesame formulation, or separate formulations.

In some embodiments, the flavivirus infection is Zika virus infection.In some embodiments, the flavivirus infection is dengue virus infection.

Optionally, the compound may be administered to a subject, or the cellcontacted, with additional agents, such as other anti-flaviviruscompounds, or other biologically active agents. For example, theadditional agent may be one or more agents useful for treating orpreventing flavivirus infection, or a symptom thereof. In someembodiments, the method further comprises administering an additionalagent, wherein the anti-flavivirus compound and the additional agent areadministered simultaneously, together within the same composition or inseparate compositions. In other embodiments, the compound and theadditional agent are administered consecutively in any order. Thus, oneor more additional agents may be administered within the sameformulation as the anti-flavivirus compound, or in a separateformulation before, during, and/or after administration of theanti-flavivirus compound.

Derivatives of any of the anti-flavivirus compounds can be synthesizedby chemical transformations of the compounds' functional groups usingstandard chemical reactions. For example, these standard chemicalreactions can include, but are not limited to: polar reactions underbasic conditions, polar reactions under acidic conditions, pericyclicreactions, and free radical reactions. In another example, thesestandard chemical reactions can include, but are not limited to:addition reactions, substitution reactions, oxidation reactions,reduction reactions, elimination reactions, hydrolysis, acylation,amidations, etherification, and esterification. Alkane functional grouptransformations can include, but are not limited to: free radicalchlorination (hv, Cl₂), free radical bromination (hv, Br₂), and allylicbromination (NBS). Alkene functional group transformations can include,but are not limited to: addition of HCl, addition of HBr, addition ofHI, addition of H₃O(+), chlorination (Cl₂) bromination (Br₂), iodination(I₂), chlorohydrin formation (Cl₂/H₂O), bromohydrin formation (Br₂/H₂O),ether formation (H⁺/ROH), oxymercuration (Hg(OAc)₂/H₂O), oxymercuration,(Hg(OAc)₂/ROH), hydroboration, epoxidation (RCO₃H), dihydroxylation(OsO₄), dihydroxylation (KMnO₄), cyclopropanation,dichlorocyclopropanation, ozonolysis (reductive workup), ozonolysis(oxidative workup), oxidative cleavage (KMnO₄), hydrogenation,rearrangements (H shift), rearrangements (alkyl shift), free radicaladdition of HBr, and Sharpless epoxidation. Alkyne functional grouptransformations can include, but are not limited to: deprotonation(acetylide formation), S_(N)2 with alkyl halides, partial reduction(Lindlar), partial, reduction (Na/NH₃), hydroboration, oxymercuration,addition of HCl, HBr, or HI, addition of HCl, HBr, or HI, hydrogenation,ozonolysis, oxidative cleavage (KMnO₄), and halogenation (Cl₂, Br₂, I₂).The substitution reaction can include, but is not limited to: alcoholformation, nitrile formation, thiol formation, ether formation,thioether formation, azides, ester formation, acetylide addition,alkanes (Gilman reagents), ammonium salt formation, alkyl chlorideformation, alkyl bromide formation, alkyl iodide formation, alkyl shift,and hydride shift. Elimination reactions can include, but are notlimited to: alkenes from alkyl halides, alkenes from alcohols (strongacid), alkenes from alcohols (POCl₃), alkenes from alkyl halides, E1with rearrangement (alkyl shift), Hoffmann elimination, and alkyneformation via elimination E1 with rearrangement (hydride shift).Organometallic reactions can include, but are not limited to: Grignardformation (alkyl halides), Grignard formation (alkenyl halides),reaction of Grignards with acids, addition of Grignards to aldehydes,addition of Grignards to ketones, addition of Grignards to esters,reaction of Grignards with CO₂, addition of Grignards to nitriles,formation of organolithium reagents, formation of Gilman reagents,S_(N)2 with Gilman reagents, addition of Gilman reagents to enones,addition of Gilman to acyl halides, Heck reaction, Suzuki reaction, andStille reaction. Reactions of epoxides can include, but are not limitedto: epoxide opening (basic conditions), epoxide opening (acidicconditions), epoxide opening (diol formation), epoxide formation (fromhalohydrins), epoxide formation (from alkenes), and Sharplessepoxidation of alkenes. Reactions of alcohols and thiols can include,but are not limited to: deprotonation (alkoxide formation), protonation(onium ion formation), conversion to tosylates/mesylates, conversion toalkyl chlorides (SOCl₂), conversion to alkyl bromides (PBr₃), oxidationto aldehydes (PCC), oxidation to ketones (PCC+others), oxidation tocarboxylic acid, (H₂CrO₄+others), protection as silyl ethers, thiolformation (S_(N)2), and thiol oxidation to disulfides. Reactions ofdienes can include, but are not limited to: Diels-alder reaction,polymerization of dienes, reactions of aromatics (arenes), nitration(HNO₃/H₂SO₄), chlorination (Cl₂ plus catalyst), bromination (Br₂ pluscatalyst), sulfonylation (SO₃/H₂SO₄), Friedel Crafts alkylation (R-Xplus catalyst), Friedel Crafts acylation (RCOX plus catalyst),iodination (I₂/catalyst), Side chain oxidation (KMnO₄), reduction ofnitro groups, reduction of aromatic ketones, Side chain bromination,nucleophilic aromatic substitution (S_(N)Ar), and aryne formation(S_(N)Ar via arynes). Reactions of aldehydes and ketones can include,but are not limited to: hydrate formation (H₂O), cyanohydrin formation(CN), reduction of aldehydes (NaBH₄), reduction of aldehydes (LiAlH₄),reduction of ketones (NaBH₄), reduction of ketones (LiAlH₄), Grignardaddition to aldehydes, Grignard addition to ketones, acetal formation(ROH/H⁺), acetal hydrolysis (H₃O⁺), imine, formation (RNH₂), Enamineformation (R₂NH), Wolff-Kishner: reduction to alkanes, Clemmensen,reduction to alkanes, oxidation to carboxylic acid (H₂CrO₄ or KMnO₄),keto-enol tautomerism, enolate formation, aldol addition reaction,alkylation of enolates, Wittig reaction (alkene formation), thioacetalformation, imine hydrolysis, oxidation to carboxylic acids (Tollens),haloform reaction, Baeyer-Villiger reaction, aldol condensation,Cannizarro reaction. Reactions of carboxylic acids can include, but arenot limited to: deprotonation (carboxylate formation), formation viaGrignard and CO₂, conversion to acid chloride (SOCl₂), reduction(LiAlH₄), Fischer esterification, and decarboxylation (of β-keto acids).Reactions of esters can include, but are not limited to: reduction toaldehydes (DIBAL-H), reduction to alcohols (LiAlH₄), hydrolysis tocarboxylic acid (acidic), hydrolysis to carboxylic acid (basic),addition of Grignard reagents to esters, Claisen condensation, andtransesterification (basic conditions). Reactions of acyl halides caninclude, but are not limited to: conversion to esters (ROH), conversionto carboxylic acids (H₂O), conversion to anhydrides (RCO₂), conversionto amides (RNH₂), conversion to ketones (Gilman reagents), andconversion to aldehydes (LiAlH(OtBu)₃). Reactions of α,β-unsaturatedketones (enones) can include, but are not limited to: Michael reaction(conjugate addition of enolates), conjugate addition of Gilman reagents,conjugate addition of other nucleophiles. Reactions of amines and amidescan include, but are not limited to: dehydration of amides to nitriles(P₂O₅), Hofmann rearrangement, Gabriel synthesis of amines, reductiveamination, formation of diazonium salts, reactions of diazonium salts,amide formation using DCC, amide formation from acid halides, andCurtius rearrangement. Reactions of nitriles can include, but are notlimited to: addition of Grignard reagents to nitriles, reduction toamines (LiAlH₄), hydrolysis to carboxylic acids. Optionally, potentialderivatives of compounds disclosed herein can be tested for the abilityto inhibit virally-induced apoptosis and/or suppress viral replication,viral protein production using methods disclosed herein or using othermethods known in the art (e.g., ATP cell viability assay).

In some embodiments of the methods, compositions, kits, and packageddosage formulations of the invention, the Flavivirus infection is Zikavirus, West Nile virus, dengue virus (e.g., type 1, 2, 3, or 4),tick-borne encephalitis virus, Japanese encephalitis virus, St. Louisencephalitis virus, or yellow fever virus. Other members of the genuscan be found in Kuno G et al., Journal of Virology, 1998, “Phylogeny ofthe Genus Flavivirus,” 72(1):73-83, which is incorporated herein byreference. In some embodiments, the Flavivirus is Zika virus. The Zikavirus may be any origin or lineage (e.g., African, Asian, American,Brazilian). Examples of Zika virus strains include but are not limitedto MR766 (1947 Uganda strain), FSS13025 (2010 Cambodian strain),PRVABC59 (2015 Puerto Rican strain), GZ01/2016 (2016 Chinese strain (exVenezuela)), H/PF/2013 (2013 French Polynesian strain), IBH30656 (1968Nigerian strain), Paraiba 2015 (2015 Brazilian strain), PLCal_ZV (2013Canadian strain (ex Thailand)), SMGC-1 (2016 Chinese strain), SPH 2015(2015 Brazilian strain), and SZ01 (2016 Chinese strain).

In each of the aforementioned methods, compositions, kits, and dosageformulations, additional agents (i.e., in addition to the one or moreanti-flavivirus compounds) may be administered to the subject, contactedwith the cell, or included in the compositions. In some embodiments, theadditional agent is useful for the treatment or prevention of flavivirusinfection, such as Zika virus infection or Dengue virus infection.

In some embodiments of the methods, compositions, kits, and packageddosage formulations of the invention, two or more compounds useful fortreating or preventing Zika virus, Dengue virus, or other flavivirusinfection by distinct mechanisms of action from one another may beutilized. For example, compounds of the invention can inhibit or reduceZika virus, Dengue virus, or other flavivirus replication, and thusrepresent one class of compounds of the invention. A combination ofcompounds may exhibit a synergistic effect in protecting cells fromflavivirus-induced (e.g., Zika virus-induced or dengue-virus induced)cell death.

The methods of the invention may be used to treat an existing flavivirusinfection in a subject, or the methods of the invention may be usedprophylactically to prevent a flavivirus infection in a subject. As usedherein, in this context, the term “prevent” or “prevention” is inclusiveof delaying the onset of infection and/or one or more symptoms ofinfection, and precluding the occurrence or reoccurrence of infectionand/or one or more symptoms of infection. Thus, in some embodiments, thesubject has the flavivirus infection at the time the at least onecompound is administered, and the at least one compound is administeredas therapy.

In some embodiments, the methods further comprise, prior toadministering the compound to the subject, identifying the subject ashaving the flavivirus infection. The identifying step may compriseassaying a biological sample (e.g., blood, saliva, or urine) obtainedfrom the subject for the presence of flavivirus nucleic acids orflavivirus proteins (e.g., Zika virus nucleic acids or Zika virusproteins, or dengue virus nucleic acids or dengue virus proteins). Insome embodiments, assaying includes the use of reversetranscriptase-polymerase chain reaction (RT-PCR), immunological assay(e.g., ELISA), or Plaque-reduction neutralization testing (PRNT).

One or more compounds of the invention (also referred to herein as theactive ingredients) are administered by any route appropriate to thecondition to be treated. Suitable routes include oral, rectal, nasal,topical (including buccal and sublingual), vaginal and parenteral(including subcutaneous, intramuscular, intravenous, intradermal,intrathecal and epidural), and the like. It will be appreciated that thepreferred route may vary with for example the condition of the subject.In some embodiments, at least one compound of the invention isadministered orally, nasally, rectally, parenterally, subcutaneously,intramuscularly, or intravascularly (e.g., intravenously).

Another aspect of the invention concerns a packaged dosage formulationcomprising at least one anti-flavivirus one compound in apharmaceutically acceptable dosage in one or more packages, packets, orcontainers.

Another aspect of the invention concerns a kit comprising, in one ormore containers, at least one anti-flavivirus compound.

In some embodiments, the kit comprises a combination of two or moreanti-flavivirus compounds of the invention. In some embodiments, the kitfurther comprises an additional agent effective for the treatment orprevention of Flavivirus virus infection. In some embodiments, the kitfurther comprises an additional agent effective for the treatment of oneor more symptoms of flavivirus infection.

Various techniques may be used to increase bioavailability of theanti-flavivirus compounds of the invention. Prodrugs employ variousphysical and chemical modifications to improve features of the activedrug, and in some embodiments may be viewed as pharmacologicallyinactive prodrug functional groups that undergo a chemicaltransformation or enzymatic cleavage to liberate the active parent drugand produce the desired effect in the body. Utilizing a prodrug approachcan yield benefits such as enhanced solubility, improved selectivetargeting of drugs to anatomical sites, protection from rapid metabolismand elimination, reduction toxic effects of an active drug on otherparts of the body, and enhanced patient compliance.

Chemical reactions, reactants, and reagents that may be utilized toenhance solubility and make prodrugs of compounds are described inMarch's Advanced Organic Chemistry, 7^(th) edition, 2013, Michael B.Smith, which is incorporated herein by reference in its entirety.

Compounds, and compositions comprising them, useful in the methods ofthe subject invention can be formulated according to known methods forpreparing pharmaceutically useful compositions. Formulations aredescribed in detail in a number of sources which are well known andreadily available to those skilled in the art. For example, Remington'sPharmaceutical Science by E. W. Martin describes formulations which canbe used in connection with the subject invention. In general, thecompositions of the subject invention will be formulated such that aneffective amount of at least one compound of the invention is combinedwith a suitable carrier or diluent in order to facilitate effectiveadministration of the composition. The compositions used in the presentmethods can also be in a variety of forms. These include, for example,solid, semi-solid, and liquid dosage forms, such as tablets, pills,powders, liquid solutions or suspension, suppositories, injectable andinfusible solutions, and sprays. The preferred form depends on theintended mode of administration and therapeutic application. Thecompositions also preferably include conventional pharmaceuticallyacceptable carriers or diluents which are known to those skilled in theart. Examples of carriers or diluents for use with the subject peptidesand polynucleotides include, but are not limited to, water, saline, oilsincluding mineral oil, ethanol, dimethyl sulfoxide, gelatin,cyclodextrans, magnesium stearate, dextrose, cellulose, sugars, calciumcarbonate, glycerol, alumina, starch, and equivalent carriers anddiluents, or mixtures of any of these. Formulations of the compounds ofthe invention can also comprise suspension agents, protectants,lubricants, buffers, preservatives, and stabilizers. To provide for theadministration of such dosages for the desired therapeutic treatment,pharmaceutical compositions of the invention will advantageouslycomprise between about 0.1% and 45%, and especially, 1 and 15% by weightof the total of one or more of the peptide or polynucleotide based onthe weight of the total composition including carrier or diluent.

The anti-flavivirus compounds of the subject invention can also beadministered utilizing liposome technology, slow release capsules,implantable pumps, and biodegradable containers. These delivery methodscan, advantageously, provide a uniform dosage over an extended period oftime.

The anti-flavivirus compounds of the invention can also be modified bythe addition of chemical groups, such as PEG (polyethylene glycol).PEGylated compounds can exhibit extended half-lives in vivo incomparison to compounds that are not PEGylated when administered invivo. Compounds can also be modified to improve cell membranepermeability. In one embodiment, cell membrane permeability can beimproved by attaching a lipophilic moiety, such as a steroid, to thecompound. In another embodiment, compounds of the invention can becoupled to a cell-penetrating peptide (CPP). CPPs are typically shortpeptides that are highly cationic and typically include several arginineand/or lysine amino acids. CPPs can be classified as hydrophilic,amphiphilic, or periodic sequence.

The subject invention also concerns a packaged dosage formulationcomprising in one or more containers at least compound, and/orcomposition of the subject invention formulated in a pharmaceuticallyacceptable dosage. The package can contain discrete quantities of thedosage formulation, such as tablet, capsules, lozenge, and powders. Thequantity of compound in a dosage formulation and that can beadministered to a patient can vary from about 1 mg to about 5000 mg, orabout 1 mg to about 2000 mg, or more typically about 1 mg to about 500mg, or about 5 mg to about 250 mg, or about 10 mg to about 100 mg.

Optionally, the methods include, prior to administration of at least onecompound of the invention, or re-administration of at least one compoundof the invention, determining whether the subject has a Flavivirusinfection (e.g., a Zika virus infection or dengue virus infection) orone or more symptoms consistent with a Flavivirus infection. Theselectivity will vary with the detection method. Some methods canidentify the infection only as a Flavivirus infection. Other methods canmore selectively detect and identify the infection as a particularFlavivirus infection, such as a dengue virus infection, or specific typeof dengue virus infection, for example.

The terms “selectively detect” or “selectively detecting” refer to thedetection of Flavivirus, nucleic acids using oligonucleotides, e.g.,primers, probes and/or capture oligonucleotides that are capable ofdetecting a particular Flavivirus virus nucleic acid, for example, byamplifying and/or binding to at least a portion of an RNA segment from aparticular type of Flavivirus, but do not amplify and/or bind tosequences from other types of Flavivirus under appropriate hybridizationconditions. For example, in the context of DENV, the terms “selectivelydetect” or “selectively detecting” refer to the detection of denguevirus nucleic acids using oligonucleotides, e.g., primers, probes and/orcapture oligonucleotides that are capable of detecting a particulardengue virus nucleic acid, for example, by amplifying and/or binding toat least a portion of an RNA segment from a particular type of denguevirus, such as a particular dengue virus serotype (e.g., DENV-1, DENV-2,DENV-3, or DENV-4), but do not amplify and/or bind to sequences fromother types of dengue viruses under appropriate hybridizationconditions.

In the case of Zika virus, during the first week after onset ofsymptoms, viral RNA can often be identified in serum; thus, Zika virusdisease can be diagnosed by performing reverse transcriptase-polymerasechain reaction (RT-PCR) on serum. Urine and saliva samples may also beused for detection of Zika virus (Gourinat A-C et al. (2015) EmergInfect Dis, vol. 21, no. 1, pp. 84-86; and Musso D et al. (2015) J ClinVirol, vol. 68, pp. 53-55).

Virus-specific IgM and neutralizing antibodies typically develop towardthe end of the first week of illness; cross-reaction with relatedflaviviruses (e.g., dengue and yellow fever viruses) is common and maybe difficult to discern. Plaque-reduction neutralization testing (PRNT)can be performed to measure virus-specific neutralizing antibodies anddiscriminate between cross-reacting antibodies in primary flavivirusinfections.

Conventional laboratory techniques/tools are available for the diagnosisof DENV. Confirmation of DENV can be made through virus isolation,genome amplification, as well, as antigen and antibody detection viaserology. Selection of a suitable test can be dependent on a variety offactors such as viremia period and infection status (primary orsecondary). Examples of tests include viral culture using biologicalsamples such as plasma, serum peripheral blood, cerebrospinal fluid,pleural fluid, and immune system tissues, such as the liver, spleen, andlymph node; nucleic acid amplification (e.g., by RT-PCR); andserological diagnosis such as hemagglutination inhibition (HI) assay,detection of dengue-specific IgM antibody (e.g., by ELISA or rapidtest), detection of dengue-specific IgG antibody, and detection ofdengue NS1 (e.g., by ELISA or rapid test). Methods and tools fordetection of DENV and diagnosis of dengue infection can be found, forexample, in Nguyen et al. (Int. J. Mol. Sci., 2019, 20:3464), Pang etal. (J. Clin. Microbiol., 2017, 55:3339-3349), Parkash et al. (Viruses,2015, 7:5410-5427), Muller et al. (J. Infect. Dis., 2017, 215(Suppl2):S89-S95), Fatima et al. (Pak. J. Pharm. Sci., 2015, 28:271-280),Huhtamo et al. (J. Clin. Virol., 2010, 47:49-53), which are eachincorporated herein by reference in their entirety.

In the case of Zika virus, some infected individuals will not know theyhave the disease because they will not have symptoms. The most commonsymptoms of Zika virus infection are fever, maculo-papular rash (oftenspreading from face to body), joint pain, retro-orbital pain, orconjunctivitis (red eyes). Other common symptoms include generalnon-specific such as myalgia, asthenia, and headache. The incubationperiod (the time from exposure to symptoms) for Zika virus disease isnot known, but is likely to be a few days to a week. The illness isusually mild with symptoms lasting for several days to a week afterbeing bitten by an infected mosquito. The Zika virus usually remains inthe blood of an infected person for approximately a week but it can befound longer in some individuals.

In the case of dengue virus (DENV), dengue fever causes a spectrum ofpresentations ranging from mild self-limiting illness to severe disease.Clinical symptoms of DENV may include one or more of the following:flu-like symptoms, rash, diarrhea, muscle/joint pain, neurologicalmanifestation, bleeding tendency, and thrombocytopenia. Subjectspresenting with all four criteria of fever, hemmorhagic diathesis,thrombocytopenia, and evidence of plasma leakage will typically beclassified as having DHF, or DSS if they present with symptoms of shock(based on 1997 WHO dengue classification). The severity classificationnow includes dengue, dengue with warning signs, and severe dengue. Thesubject may have any of these before or after administration of the atleast one compound.

Treatment methods optionally include steps of advising that the subjectget plenty of rest and drink fluids for hydration and administration ofagents that alleviate symptoms of Flavivirus infection (e.g., denguevirus infection or Zika virus infection), such as those that reducefever and pain (e.g., acetaminophen and/or paracetamol). The methods mayinclude administration of the fluids to the subject for hydration.

The subject may be any age or gender. In some embodiments, the subjectis male. In some embodiments, the subject is female. In someembodiments, the subject is a post-pubescent female. In someembodiments, the subject is a post-pubescent, pre-menopausal female. Insome embodiments, the subject is a non-pregnant female. In someembodiments, the subject is a pregnant female. In some embodiments, thesubject has Guillain-Barré syndrome or another condition that isassociated with ZIKV infection.

In some embodiments, the Flavivirus is DENV, the subject has mild orsevere dengue fever at the time of administration, and at least onecompound is administered to the subject as therapy. In some embodiments,the Flavivirus is DENV, at least one compound is administered prior toinfection, and administration prevents or delays onset of dengue fever(mild or severe dengue fever). In some embodiments, the at least onecompound is administered to the subject as therapy for denguehemorrhagic fever (DHF) or dengue shock syndrome (DSS), or to prevent ordelay the onset of DHF or DSS.

The invention further provides kits, including at least one compound ofthe invention and pharmaceutical formulations, packaged into suitablepackaging material, optionally in combination with instructions forusing the kit components, e.g., instructions for performing a method ofthe invention. In one embodiment, a kit includes an amount of at leastone compound of the invention, and instructions for administering atleast one compound of the invention to a subject in need of treatment ona label or packaging insert. In further embodiments, a kit includes anarticle of manufacture, for delivering at least one compound of theinvention into a subject locally, regionally or systemically, forexample.

As used herein, the term “packaging material” refers to a physicalstructure housing the components of the kit. The packaging material canmaintain the components in a sterile state, and can be made of materialcommonly used for such purposes (e.g., paper, corrugated fiber, glass,plastic, foil, ampules, etc.). The label or packaging insert can includeappropriate printed and/or digital instructions, for example, forpracticing a method of the invention, e.g., treating a Flavivirusinfection (e.g., Zika virus infection or dengue virus infection), anassay for identifying a subject having a Flavivirus infection (e.g.,Zika virus infection or dengue virus infection), etc. Thus, inadditional embodiments, a kit includes a label or packaging insertincluding instructions for practicing a method of the invention insolution, in vitro, in vivo, or ex vivo.

Instructions can therefore include instructions for practicing any ofthe methods of the invention described herein. For example,pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration to a subject totreat a flavivirus infection (e.g., Zika virus infection or dengue virusinfection). Instructions may additionally include indications of asatisfactory clinical endpoint or any adverse symptoms that may occur,storage information, expiration date, or any information required byregulatory agencies such as the Food and Drug Administration or EuropeanMedicines Agency for use in a human subject.

The instructions may be digital or on “printed matter,” e.g., on paperor cardboard within the kit, on a label affixed to the kit or packagingmaterial, or attached to a vial or tube containing a component of thekit. Instructions may comprise voice or video tape and additionally beincluded on a computer readable medium, such as a disk (diskette or harddisk), optical CD such as CD- or DVD-ROM/RAM, magnetic tape, electricalstorage media such as RAM and ROM and hybrids of these such asmagnetic/optical storage media.

Kits can additionally include a buffering agent, a preservative, or anagent for stabilizing at least one compound of the invention. The kitcan also include components for assaying for the presence of Zika virus,dengue virus, or other Flavivirus, e.g., an antibody or antibodyfragment specific for a Zika virus, dengue virus, or other Flavivirusantigen, one or more primers specific for Zika virus, dengue virus, orother Flavivirus nucleic acids, a control sample or a standard. Eachcomponent of the kit can be enclosed within an individual container orin a mixture and all of the various containers can be within single ormultiple packages.

Kits can include packaging material that is compartmentalized to receiveone or more containers such as vials, tubes, and the like, each of thecontainer(s) including one of the separate elements to be used in amethod described herein. Packaging materials for use in packagingpharmaceutical products include, by way of example only U.S. Pat. Nos.5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, pumps, bags, vials, light-tight sealed containers, syringes,bottles, and any packaging material suitable for a selected formulationand intended mode of administration and treatment.

A kit may include one or more additional containers, each with one ormore of various materials desirable from a commercial and userstandpoint for use of the compounds for treating or preventing Zikavirus, dengue virus, or other Flavivirus infection. Non-limitingexamples of such materials include, but not limited to, buffers,diluents, carrier, package, container, vial and/or tube labels listingcontents and/or instructions for use, and package inserts withinstructions for use.

A label can be on or associated with a container containing a compoundof the invention. A label can be on a container when letters, numbers orother characters forming the label are attached, molded or etched intothe container itself; a label can be associated with a container when itis present within a receptacle or carrier that also holds the container,e.g., as a package insert. A label can be used to indicate that thecontents are to be used for a specific therapeutic application. Thelabel can also indicate directions for use of the contents, such as inthe methods described herein.

In some embodiments of the kit, the compound(s) of the invention can bepresented in a pack or dispenser device which can contain one or moreunit dosage forms containing a compound disclosed herein. The pack canfor example contain metal or plastic foil, such as a blister pack. Thepack or dispenser device can be accompanied by instructions foradministration. The pack or dispenser can also be accompanied with anotice associated with the container in form prescribed by agovernmental agency regulating the manufacture, use, or sale ofpharmaceuticals, which notice is reflective of approval by the agency ofthe form of the drug for human or veterinary administration. Suchnotice, for example, can be the labeling approved by the U.S. Food andDrug Administration for prescription drugs, or the approved productinsert. Compositions containing a compound provided herein formulated ina compatible pharmaceutical carrier can also be prepared, placed in anappropriate container, and labeled for treatment of an indicatedcondition.

Additional anti-flavivirus compounds can be identified by determiningwhether the candidate compounds reduce virally-induced apoptosis and/orsuppress viral replication.

DEFINITIONS

The terms “compounds of the invention”, “compounds of the presentinvention”, “anti-flavivirus compound of the invention”, or“anti-flavivirus compound of the invention” (unless specificallyidentified otherwise), and grammatical variations thereof, refer to thecompounds and classes of compounds disclosed herein, such as at leastone compound selected from the group consisting of: NCGC00102779-01,NCGC00113159-01, NCGC00104879-01, NCGC00025125-18, NCGC00018238-09,NCGC00112058-01, NCGC00169957-03, NCGC00387651-01, NCGC00179895-03,NCGC00108525-01, NCGC00071621-03, NCGC00131231-01, NCGC00246910-02,NCGC00108581-01, NCGC00107055-01, NCGC00263862-02, NCGC00072088-02,NCGC00357393-02, and NCGC00378623-01 (the chemical structures of whichare shown in Table 1), or a prodrug, metabolite, or derivative of any ofthe foregoing, or a pharmaceutically acceptable salt of any of theforegoing. Compounds of the invention include stereoisomers (includingdiastereoisomers and enantiomers), rotamers, tautomers and isotopicallylabeled compounds (including deuterium substitutions), as well asinherently formed moieties (e.g., polymorphs, solvates and/or hydrates)of the aforementioned compounds. For purposes of this invention,solvates and hydrates are generally considered compositions.

The term “a,” “an,” “the” and similar terms used in the context of thepresent invention (especially in the context of the claims) are to beconstrued to cover both the singular and plural unless otherwiseindicated herein or clearly contradicted by the context. Thus, forexample, reference “a cell” or “a compound” should be construed to coverboth a singular cell or singular compound and a plurality of cells and aplurality of compounds unless indicated otherwise or clearlycontradicted by the context.

The term “agent” refers to all materials that may be used as or in apharmaceutical composition, or that may be a compound such as smallsynthetic or naturally derived organic compounds, nucleic acids,polypeptides, antibodies, fragments, isoforms, variants, or othermaterials that may be used independently for such purposes.

The term “small molecule” refers to a composition that has a molecularweight of less than about 3 kilodaltons (kDa), less than about 1 kDa, orless than about 1 kDa. Small molecules may be nucleic acids, peptides,polypeptides, peptidomimetics, carbohydrates, lipids, or other organic(carbon-containing) or inorganic molecules. A “small organic molecule”is an organic compound (or organic compound complexed with an inorganiccompound (e.g., metal), that has a molecular weight of less than about 3kDa, less than about 1.5 kDa, or less than about 1 kDa.

The term “isolated,” when used as a modifier of a composition of matter,such as a compound, means that the compositions are made by the hand ofman or are separated from their naturally occurring in vivo environment.Generally, compositions so separated are substantially free of one ormore materials with which they normally associate with in nature, forexample, one or more protein, nucleic acid, lipid, carbohydrate, cellmembrane. A “substantially pure” molecule can be combined with one ormore other molecules. Thus, the term “substantially pure” does notexclude combinations of compositions. Substantial purity can be at leastabout 60% or more of the molecule by mass. Purity can also be about 70%or 80% or more, and can be greater, for example, 90% or more. Purity canbe determined by any appropriate method, including, for example, UVspectroscopy, chromatography (e.g., HPLC, gas phase), gelelectrophoresis (e.g., silver or coomassie staining) and sequenceanalysis (for nucleic acid and peptide). The compounds of the inventionmay be in isolated or substantially pure form.

The present invention includes derivatives of identified compounds, alsoreferred to herein as pharmaceutically active derivatives.“Pharmaceutically active derivative” refers to any compound that uponadministration to the subject or cell, is capable of providing directlyor indirectly, the activity disclosed herein. The term “indirectly” alsoencompasses prodrugs which may be converted to the active form of thedrug via endogenous enzymes or metabolism. The prodrug is a derivativeof the compounds according to the invention and presenting flavivirus(e.g., Zika virus or dengue virus) inhibitory activity and/or protectiveactivity against effects of flavivirus (e.g., Zika virus or denguevirus) that has a chemically or metabolically decomposable group, and acompound that may be converted into a pharmaceutically active compoundaccording to the invention in vivo by solvolysis under physiologicalconditions. The prodrug is converted into a compound according to thepresent invention by a reaction with an enzyme, gastric acid or the likeunder a physiological condition in the living body, e.g., by oxidation,reduction, hydrolysis or the like, each of which is carried outenzymatically. These compounds can be produced from compounds of theinvention according to well-known methods. The term “indirectly” alsoencompasses metabolites of compounds according to the invention.Chemical reactions, reactants, and reagents useful for makingderivatives can be found, for example, in March's Advanced OrganicChemistry, 7^(th) edition, 2013, Michael B. Smith, which is incorporatedherein by reference in its entirety.

The term “metabolite” refers to all molecules derived from any of thecompounds according to the invention in a cell or organism, preferablymammal. Pharmaceutically active metabolites of the compounds of theinvention may be administered to a subject or contacted with a cell invitro or in vivo.

The term “prodrug” refers to a chemical compound that can be convertedby the body (i.e., biotransformed) to another chemical compound that haspharmacological activity. The prodrug may itself have pharmacologicalactivity before conversion, or be inactive before conversion andactivated upon conversion. Active prodrugs or inactive prodrugs ofcompounds of the invention may be administered to a subject or contactedwith a cell in vitro or in vivo. Instead of administering a drugdirectly, a prodrug may be used instead to improve how a drug isabsorbed, distributed, metabolized, and excreted (ADME). For example, aprodrug may be used to improve bioavailability when a drug itself ispoorly absorbed from the gastrointestinal tract, or to improve howselectively the drug interacts with cells or processes that are not itsintended target, which can reduce adverse or unintended effects of adrug. Major types of prodrugs include, but are not limited to, type Iprodrugs, which are biotransformed inside cells (intracellularly), andtype II prodrugs, which are biotransformed outside cells(extracellularly), such as in digestive fluids or in the body'scirculatory system. These types can be further categorized into subtypesbased on factors such as whether the intracellular bioactivationlocation is also a site of therapeutic action, or whether or notbioactivation occurs in the gastrointestinal fluids or in thecirculation system (Wu, Kuei-Meng, “A New Classification of Prodrugs:Regulatory Perspectives, Pharmaceuticals, 2009, 2(3):77-81, which isincorporated by reference herein in its entirety).

Pharmaceutical formulations include “pharmaceutically acceptable” and“physiologically acceptable” carriers, diluents or excipients. As usedherein the terms “pharmaceutically acceptable” and “physiologicallyacceptable” carriers, diluents, or excipients include solvents (aqueousor non-aqueous), solutions, emulsions, dispersion media, coatings,isotonic and absorption promoting or delaying agents, compatible withpharmaceutical administration. Such formulations can be contained in aliquid; emulsion, suspension, syrup or elixir, or solid form; tablet(coated or uncoated), capsule (hard or soft), powder, granule, crystal,or micro-bead. Supplementary compounds (e.g., preservatives,antibacterial, antiviral and antifungal agents) can also be incorporatedinto the compositions. The phrase “pharmaceutically acceptable”indicates that the substance or composition must be compatiblechemically and/or toxicologically, with the other ingredients comprisinga formulation, and/or the mammal and/or cells being treated therewith.Examples of pharmaceutically acceptable carriers include but are notlimited to saline, buffered saline, isotonic saline, Ringer's solution,dextrose, sterile water, deionized water, glycerol, ethanol, 5% dextrosein water, propylene glycol, and combinations of two or more of theforegoing.

The phrase “effective amount”, in the context of a subject, means anamount of at least one compound of the invention that (i) treats orprevents the particular disease, condition, or disorder (e.g., Zika,dengue, or other flavivirus infection) in a subject, (ii) attenuates,ameliorates, or eliminates one or more symptoms of the particulardisease, condition, or disorder (e.g., Zika, dengue, or other flavivirusinfection) in a subject, or (iii) prevents or delays the onset of one ormore symptoms of the particular disease, condition, or disorderdescribed herein (e.g., Zika, dengue or other flavivirus infection) in asubject.

The phrase “effective amount”, in the context of a cell in vitro or invivo, means an amount of at least one compound of the invention that (i)treats or prevents the particular disease, condition, or disorder (e.g.,Zika, dengue, or other flavivirus infection) in a cell, (ii) attenuates,ameliorates, or eliminates one or more effects of the particulardisease, condition, or disorder (e.g., Zika, dengue, or other flavivirusinfection) in a cell, or (iii) prevents or delays the onset of one ormore effects of the particular disease, condition, or disorder describedherein (e.g., Zika, dengue, or other flavivirus infection) in a subject.

As used herein, a subject is “in need of” a treatment if such human ornon-human animal subject would benefit biologically, medically or inquality of life from such treatment (preferably, a human). In someembodiments, the subject has a flavivirus infection and is in need oftherapy. In other embodiments, the subject does not have a flavivirusinfection and is in need of prophylaxis. In some embodiments, thesubject in need of prophylaxis is at risk of becoming infected with theflavivirus. In some embodiments, the subject is at increased risk ofbecoming infected with the flavivirus relative to others in thepopulation. In some embodiments, the subject is suspected to have aflavivirus infection.

As used herein, the term “inhibit”, “inhibition” or “inhibiting” refersto the reduction or suppression of a given condition, symptom, ordisorder, or disease (e.g., Zika virus, dengue virus, or otherflavivirus infection, or Zika viral, dengue viral, or other flavivirusload or titer), or a significant decrease in the baseline activity of abiological activity or process (inhibits or suppresses Zika, dengue, orother flavivirus infection, or inhibits or suppresses Zika, dengue, orother flavivirus replication, or inhibits or suppresses Zika-induced orother flavivirus-induced neural cell death.

As used herein, the terms “subject”, “patient”, and “individual” referto a human or non-human animal. Typically, the animal is a mammal. Asubject also refers to for example, primates (e.g., humans), cows,sheep, goats, horses, dogs, cats, rabbits, rats, mice, fish, birds andthe like. In certain embodiments, the subject is a primate. In yet otherembodiments, the subject is a human. The subject may be any age orgender. For example, in some embodiments, the subject is a female. Insome embodiments, the subject is a post-pubescent female or apost-pubescent female. In some embodiments, the subject is a pregnantfemale; in other embodiments, the subject is a non-pregnant female.

In some embodiments, the subject has been exposed to the Flavivirusand/or has the Flavivirus infection at the time the at least onecompound is administered. In some embodiments, the Flavivirus is DENV,the subject has mild or severe dengue fever at the time ofadministration, and administration is provided to the subject astherapy. In some embodiments, the Flavivirus is DENV, and the at leastone compound is administered prior to infection, and administrationprevents or delays onset of dengue fever (mild or severe dengue fever).In some embodiments, the at least one compound is administered to thesubject as therapy for dengue hemorrhagic fever (DHF) or dengue shocksyndrome (DSS), or to prevent or delay the onset of DHF or DSS.

As used herein, the term “treat”, “treating” or “treatment” of anydisease or disorder refers in one embodiment, to ameliorating thedisease or disorder (i.e., slowing or arresting or reducing thedevelopment of the disease or at least one of the clinical symptomsthereof). In another embodiment “treat”, “treating” or “treatment”refers to alleviating or ameliorating at least one physical parameterincluding those which may not be discernible by the subject. In yetanother embodiment, “treat”, “treating” or “treatment” refers tomodulating the disease or disorder, either physically, (e.g.,stabilization of a discernible symptom), physiologically, (e.g.,stabilization of a physical parameter), or both. In yet anotherembodiment, “treat”, “treating” or “treatment” refers to prophylaxis(preventing or delaying the onset or development or progression of thedisease or disorder).

As used herein, the term “administration” is intended to include, but isnot limited to, the following delivery methods: topical, oral,parenteral, subcutaneous, transdermal, transbuccal, intravascular (e.g.,intravenous or intra-arterial), intramuscular, subcutaneous, intranasal,and intra-ocular administration. Administration can be local at aparticular anatomical site, such as a site of infection, or systemic.Administration can be by any route effective to treat, prevent, or delayonset of the Flavivirus infection in the subject.

As used herein, the term “contacting” in the context of contacting acell with at least one compound of the invention in vitro or in vivomeans bringing at least one compound into contact with the cell, orvice-versa, or any other manner of causing the compound and the cell tocome into contact. In those embodiments of the method for inhibitingflavivirus infection in human or non-human animal cells in vitro or invivo, when a cell is contacted with a compound in vivo, the compound isadministered to a subject, and the administration may occur by any route(e.g., topical, oral, parenteral, subcutaneous, transdermal,transbuccal, intravascular (e.g., intravenous or intra-arterial),intramuscular, subcutaneous, intranasal, and intra-ocularadministration).

The compounds of the present invention can be formulated intopharmaceutically-acceptable salt forms. Pharmaceutically-acceptablesalts of the compounds of the invention can be prepared usingconventional techniques. “Pharmaceutically acceptable salt” includesboth acid and base addition salts. A pharmaceutically acceptable salt ofany one of the compounds described herein is intended to encompass anyand all pharmaceutically suitable salt forms. Preferred pharmaceuticallyacceptable salts described herein are pharmaceutically acceptable acidaddition salts and pharmaceutically acceptable base addition salts.“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid,hydrofluoric acid, phosphorous acid, and the like. Also included aresalts that are formed with organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, alkanedioic acids, aromatic acids, aliphatic and. aromaticsulfonic acids, etc. and include, for example, acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Exemplary salts thus include sulfates,pyrosulfates, bisulfates, sulfites, bi sulfites, nitrates, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates,trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates,mandelates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,phenylacetates, citrates, lactates, malates, tartrates,methanesulfonates, and the like. Also contemplated are salts of aminoacids, such as arginates, gluconates, and galacturonates (see, forexample, Berge S. M. et al., “Pharmaceutical Salts,” Journal ofPharmaceutical Science, 66:1-19 (1997), which is hereby incorporated byreference in its entirety). Acid addition salts of basic compounds maybe prepared by contacting the free base forms with a sufficient amountof the desired acid to produce the salt according to methods andtechniques with which a skilled artisan is familiar.

“Pharmaceutically acceptable base addition salt” refers to those saltsthat retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Pharmaceutically acceptable base addition salts may beformed with metals or amines, such as alkali and alkaline earth metalsor organic amines. Salts derived from inorganic bases include, but arenot limited to, sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Salts derived from organic bases include, but are not limited to, saltsof primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, for example, isopropylamine, trimethylamine,diethylamine, triethylamine, tripropylamine, ethanolamine,diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline,betaine, ethylenediamine, ethylenedianiline, N-methylglucamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like. See Bergeet al., supra.

All patents, patent applications, provisional applications, andpublications referred to or cited herein are incorporated by referencein their entirety, including all figures and tables, to the extent theyare not inconsistent with the explicit teachings of this specification.

Following are examples that illustrate procedures for practicing theinvention. These examples should not be construed as limiting. Allpercentages are by weight and all solvent mixture proportions are byvolume unless otherwise noted.

Example 1—Antiviral Activity of Compounds Against Zika Virus and DengueVirus

The inventors have identified 19 compounds with antiviral activityagainst Zika virus and/or dengue virus. These compounds are:NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01. The compoundswere initially identified via a computational model.

These 19 compounds were further validated for antiviral activity by aZika viral production assay and also by analyzing protein production viaWestern blot in dengue virus-infected cells. Human glioblastoma cellswere treated with individual compounds for 1 hour prior to infectionwith Zika virus or dengue virus for 24 hours. After 24 hours, cellularlysate was harvested for Western blot analysis of viral protein levels(DENV infection), or supernatant collected and used to infect Vero cellsin an infectious viral titer assay (ZIKV infection). The results areshown in Table 2, below.

TABLE 2 Anti- DENV Anti-DENV Activity Activity at 1 uM at 10 uMAnti-ZIKV (SNB-19 (SNB-19 activity Sample ID and IUPAC Name cells)cells) IC50  1 NCGC00179895-03 Yes Yes 3.49E−09((2S,2′R,3′R,4′S,5′S,5a′R,9a′R)-4′-acetoxy-3′-hydroxy-5′,8′-dimethyl-2′,3′,4′,5′,7′,9a′-hexahydrospiro[oxirane-2,10′[2,5]methanobenzo[b]oxepin]-5a′(6′H)-yl)methyl acetate  2NCGC00387651-01 Yes Yes 4.38E−09(2S,2′R,3′R,4′S,5′S,5a′R,7′S,9a′R)-4′-acetoxy-5a′-(acetoxymethyl)-3′-hydroxy-5′,8′-dimethyl-2′,3′,4′,5′,5a′,6′,7′,9a′-octahydrospiro[oxirane-2,10′-[2,5]methanobenzo[b]oxepin]-7′-yl 3-methylbutanoate  3 NCGC00169957-03No Yes 6.13E−07 3-(sec-butyl)-16-isobutyl-6-isopropyl-5,8,9-trimethyldodecahydropyrrolo[1,2-d][1]oxa[4,7,10,13,16]pentaazacyclononadecine-1,4,7,10,14,17(11H,16H)-hexaone  4 NCGC00108525-01 No Yes 1.37E−07 ethyl4-(3-(4-methoxyphenyl)-6-oxo-4-(p-tolyl)-4,6-dihydropyrrolo[3,4-c]pyrazol-5(1H)-yl)benzoate  5 NCGC00246910-02 No Yes2.70E−07 7-(3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-1,7-dihydro-4H-pyrrolo[2,3-d]pyrimidin-4-one  6 NCGC00072088-02 NoYes 2.35E−06 N-(pyridin-3-yl)thiophene-2-carboxamide  7 NCGC00108581-01No Yes 2.42E−06 ethyl 4-(3-(furan-2-yl)-6-oxo-4-(p-tolyl)-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)benzoate  8 NCGC00357393-02 YesYes 6.74E−10 (2S,3R,4S,4aR)-2,3,4,7-tetrahydroxy-3,4,4a,5-tetrahydro-[1,3]dioxolo[4,5-j]phenanthridin-6(2H)-one  9 NCGC00102779-01 No No9.24E−06 5-(4-ethoxyphenyl)-4-(3-fluorophenyl)-3-thiophen-2-yl-2,4-dihydropyrrolo[3,4-c]pyrazol-6-one 10 NCGC00113159-01 No No >30 μMmethyl 4-[[5-methyl-2-(4-methylphenyl)-1,3-oxazol-4-yl]methyl]furo[3,2-b]pyrrole-5-carboxylate 11 NCGC00104879-01 No No >30μM N-[(2-chlorophenyl)methyl]-3-(2-methoxyethyl)-2,4-dioxo-1H-quinazoline-7-carboxamide 12 NCGC00025125-18 No No 7.09E−10N-[(7S)-1,2,3,10-tetramethoxy-9-oxo-6,7-dihydro-5H-benzo[a]heptalen-7-yl]acetamide 13 NCGC00018238-09 No No 2.33E−10 methylN-(6-propoxy-1H-benzimidazol-2-yl)carbamate 14 NCGC00112058-01 No No >30μM N-[(2-chlorophenypmethyl]-6,7,10-trimethylbenzo[b][1,4]benzothiazepine-3-carboxamide 15 NCGC00071621-03No No >30 μM 4-[3-(2-methoxyphenyl)-[1,2,4]triazolo[3,4-b][1,3,4]thiadiazol-6-yl]-N,N-dimethylaniline 16 NCGC00131231-01 No No7.60E−06 4-tert-butyl-N-[1-(thiophene-2-carbonyl)-3,4-dihydro-2H-quinolin-6-yl]benzamide 17 NCGC00107055-01 No No 2.85E−089-(3-bromo-4,5-dimethoxyphenyl)-3,6,8,9-tetrahydro-2H-[1,4]dioxino[2,3-g]quinolin-7-one 18 NCGC00263862-02 No No 1.65E−085-fluoro-1-((4R,5R)-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidine- 2,4(1H,3H)-dione 19NCGC00378623-01 No No 1.78E−15(S)-2-((S)-2-(dimethylamino)-3-methylbutanamido)-N-((3R,4S,5S)-3-methoxy-1-((S)-2-((S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-(((S)-2-phenyl-1-(thiazol-2-yl)ethyl)amino)propyl)pyrrolidin-1-yl)-5-methyl-1-oxoheptan-4-yl)-N,3-dimethylbutanamide

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication and the scope of the appended claims. In addition, anyelements or limitations of any invention or embodiment thereof disclosedherein can be combined with any and/or all other elements or limitations(individually or in any combination) or any other invention orembodiment thereof disclosed herein, and all such combinations arecontemplated with the scope of the invention without limitation thereto.

Exemplified Embodiments

Examples of claimed embodiments of the invention include, but are notlimited to:

Embodiment 1. A method for treating or preventing Flavivirus infectionin a human or non-human animal subject, said method comprisingadministering an effective amount of at least one compound to a subjectin need thereof, wherein the compound is selected from the groupconsisting of NCGC00102779-01, NCGC00113159-01, NCGC00104879-01,NCGC00025125-18, NCGC00018238-09, NCGC00112058-01, NCGC00169957-03,NCGC00387651-01, NCGC00179895-03, NCGC00108525-01, NCGC00071621-03,NCGC00131231-01, NCGC00246910-02, NCGC00108581-01, NCGC00107055-01,NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01,or a prodrug, metabolite, or derivative of any of the foregoing, or apharmaceutically acceptable salt of any of the foregoing.

Embodiment 2. The method of embodiment 1, wherein the Flavivirusinfection is Zika virus infection.

Embodiment 3. The method of embodiment 1, wherein the Flavivirusinfection is dengue virus infection.

Embodiment 4. The method of any one of embodiments 1 to 3, wherein thesubject has the Flavivirus infection at the time of said administering,and the at least one compound is administered as therapy.

Embodiment 5. The method of embodiment 4, further comprising, prior tosaid administering, identifying the subject as having the Flavivirusinfection.

Embodiment 6. The method of embodiment 5, wherein said identifyingcomprises assaying a biological sample obtained from the subject for thepresence of Flavivirus nucleic acids or Flavivirus proteins.

Embodiment 7. The method of embodiment 6, wherein said assayingcomprises use of reverse transcriptase-polymerase chain reaction(RT-PCR), immunological assay, or Plaque-reduction neutralizationtesting (PRNT).

Embodiment 8. The method of embodiment 1, wherein the subject does nothave the Flavivirus infection at the time of said administering, and theat least one compound is administered as prophylaxis.

Embodiment 9. The method of any preceding embodiment, wherein the atleast one compound is administered orally, nasally, rectally,parenterally, subcutaneously, intramuscularly, or intravascularly.

Embodiment 10. The method of embodiment 1, further comprisingadministering an additional agent for treating or preventing Flavivirusinfection, or a symptom thereof, in the same formulation as the at leastone compound, or in a separate formulation before, during, or afteradministration of the at least one compound.

Embodiment 11. The method of any preceding embodiment, wherein two ormore compounds selected from the group consisting of NCGC00102779-01,NCGC00113159-01, NCGC00104879-01, NCGC00025125-18, NCGC00018238-09,NCGC00112058-01, NCGC00169957-03, NCGC00387651-01, NCGC00179895-03,NCGC00108525-01, NCGC00071621-03, NCGC00131231-01, NCGC00246910-02,NCGC00108581-01, NCGC00107055-01, NCGC00263862-02, NCGC00072088-02,NCGC00357393-02, and NCGC00378623-01, or a prodrug, metabolite, orderivative of any of the foregoing, or a pharmaceutically acceptablesalt of any of the foregoing, are administered to the subject, in thesame composition or in different compositions.

Embodiment 12. The method of any preceding embodiment, wherein the atleast one compound is administered to the subject in a pharmaceuticalcomposition comprising the at least one compound and a pharmaceuticallyacceptable carrier or diluent.

Embodiment 13. A method for inhibiting flavivirus infection in human ornon-human animal cells in vitro or in vivo, said method comprisingcontacting an effective amount of at least one compound to a human ornon-human animal cell in vitro or in vivo before or after exposure ofthe cell to Flavivirus, wherein the at least one compound is selectedfrom the group consisting of NCGC00102779-01, NCGC00113159-01,NCGC00104879-01, NCGC00025125-18, NCGC00018238-09, NCGC00112058-01,NCGC00169957-03, NCGC00387651-01, NCGC00179895-03, NCGC00108525-01,NCGC00071621-03, NCGC00131231-01, NCGC00246910-02, NCGC00108581-01,NCGC00107055-01, NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, andNCGC00378623-01, or a prodrug, metabolite, or derivative of any of theforegoing, or a pharmaceutically acceptable salt of any of theforegoing.

Embodiment 14. The method of embodiment 13, wherein the Flavivirusinfection is Zika virus infection.

Embodiment 15. The method of embodiment 13, wherein the Flavivirusinfection is dengue virus infection.

Embodiment 16. A packaged dosage formulation comprising at least onecompound, wherein the at least one compound is in a pharmaceuticallyacceptable dosage in one or more packages, packets, or containers, andwherein the at least one compound is selected from the group consistingof NCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing.

Embodiment 17. The packaged dosage formulation of embodiment 16,comprising two or more of the compounds selected from the groupconsisting of NCGC00102779-01, NCGC00113159-01, NCGC00104879-01,NCGC00025125-18, NCGC00018238-09, NCGC00112058-01, NCGC00169957-03,NCGC00387651-01, NCGC00179895-03, NCGC00108525-01, NCGC00071621-03,NCGC00131231-01, NCGC00246910-02, NCGC00108581-01, NCGC00107055-01,NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01,or a prodrug, metabolite, or derivative of any of the foregoing, or apharmaceutically acceptable salt of any of the foregoing.

Embodiment 18. A kit comprising, in one or more containers, at least onecompound; and packaging material, wherein the at least one compound isselected from the group consisting of NCGC00102779-01, NCGC00113159-01,NCGC00104879-01, NCGC00025125-18, NCGC00018238-09, NCGC00112058-01,NCGC00169957-03, NCGC00387651-01, NCGC00179895-03, NCGC00108525-01,NCGC00071621-03, NCGC00131231-01, NCGC00246910-02, NCGC00108581-01,NCGC00107055-01, NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, andNCGC00378623-01, or a prodrug, metabolite, or derivative of any of theforegoing, or a pharmaceutically acceptable salt of any of theforegoing.

Embodiment 19. The kit of embodiment 18, further comprising instructionsfor using the at least one compound or composition (e.g., for treatingor preventing a flavivirus infection, such as Zika virus infection ordengue virus infection).

Embodiment 20. The kit of embodiment 18 or 19, comprising two or more ofthe compounds selected from the group consisting of NCGC00102779-01,NCGC00113159-01, NCGC00104879-01, NCGC00025125-18, NCGC00018238-09,NCGC00112058-01, NCGC00169957-03, NCGC00387651-01, NCGC00179895-03,NCGC00108525-01, NCGC00071621-03, NCGC00131231-01, NCGC00246910-02,NCGC00108581-01, NCGC00107055-01, NCGC00263862-02, NCGC00072088-02,NCGC00357393-02, and NCGC00378623-01, or a prodrug, metabolite, orderivative of any of the foregoing, or a pharmaceutically acceptablesalt of any of the foregoing.

REFERENCES

-   1. Dick, G. W., Kitchen, S. F. & Haddow, A. J. Zika virus. I.    Isolations and serological specificity. Trans R Soc Trop Med Hyg 46,    509-520 (1952).-   2. Duffy, M. R., et al. Zika virus outbreak on Yap Island, Federated    States of Micronesia. N Engl J Med 360, 2536-2543 (2009).-   3. Cao-Lormeau, V. M., et al. Zika virus, French polynesia, South    pacific, 2013. Emerg Infect Dis 20, 1085-1086 (2014).-   4. Musso, D. Zika Virus Transmission from French Polynesia to    Brazil. Emerg Infect Dis 21, 1887 (2015).-   5. Heymann, D. L., et al. Zika virus and microcephaly: why is this    situation a PHEIC? Lancet 387, 719-721 (2016).-   6. Mlakar, J., et al. Zika Virus Associated with Microcephaly. N    Engl J Med 374, 951-958 (2016).-   7. Rasmussen, S. A., Jamieson, D. J., Honein, M. A. &    Petersen, L. R. Zika Virus and Birth Defects—Reviewing the Evidence    for Causality. N Engl J Med 374, 1981-1987 (2016).-   8. Cao-Lormeau, V. M., et al. Guillain-Barre Syndrome outbreak    associated with Zika virus infection in French Polynesia: a    case-control study. Lancet 387, 1531-1539 (2016).-   9. Araujo, L. M., Ferreira, M. L. & Nascimento, O. J. Guillain-Barre    syndrome associated with the Zika virus outbreak in Brazil. Arq    Neuropsiquiatr 74, 253-255 (2016).-   Nguyen et al. (Int. J. Mol. Sci., 2019, 20:3464)-   Pang et al. (J. Clin. Microbiol., 2017, 55:3339-3349)-   Mustafa et al. (Med. J. Armed Forces India, 2015, 71:67-70)-   Parkash et al. (Viruses, 2015, 7:5410-5427)-   Muller et al. (J. Infect. Dis., 2017, 215(Suppl 2):589-595)-   Fatima et al. (Pak. J. Pharm. Sci., 2015, 28:271-280)-   Huhtamo et al. (J. Clin. Virol., 2010, 47:49-53)

We claim:
 1. A method for treating or preventing Flavivirus infection ina human or non-human animal subject, said method comprisingadministering an effective amount of at least one compound to a subjectin need thereof, wherein the compound is selected from the groupconsisting of NCGC00102779-01, NCGC00113159-01, NCGC00104879-01,NCGC00025125-18, NCGC00018238-09, NCGC00112058-01, NCGC00169957-03,NCGC00387651-01, NCGC00179895-03, NCGC00108525-01, NCGC00071621-03,NCGC00131231-01, NCGC00246910-02, NCGC00108581-01, NCGC00107055-01,NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01,or a prodrug, metabolite, or derivative of any of the foregoing, or apharmaceutically acceptable salt of any of the foregoing.
 2. The methodof claim 1, wherein the Flavivirus infection is dengue virus infection.3. The method of claim 1, wherein the subject has the Flavivirusinfection at the time of said administering, and the at least onecompound is administered as therapy.
 4. The method of claim 1, whereinthe Flavivirus infection is dengue virus infection, and wherein thesubject has the dengue virus infection at the time of saidadministering, and the at least one compound is administered as therapy.5. The method of claim 3, further comprising, prior to saidadministering, identifying the subject as having the Flavivirusinfection.
 6. The method of claim 5, wherein said identifying comprisesassaying a biological sample obtained from the subject for the presenceof Flavivirus nucleic acids or Flavivirus proteins.
 7. The method ofclaim 6, wherein said assaying comprises use of reversetranscriptase-polymerase chain reaction (RT-PCR), immunological assay,or Plaque-reduction neutralization testing (PRNT).
 8. The method ofclaim 1, wherein the subject does not have the Flavivirus infection atthe time of said administering, and the at least one compound isadministered as prophylaxis.
 9. The method of claim 1, wherein the atleast one compound is administered to the subject orally, nasally,rectally, parenterally, subcutaneously, intramuscularly, orintravascularly.
 10. The method of claim 1, further comprisingadministering an additional agent to the subject, for treating orpreventing Flavivirus infection, or a symptom thereof, in the sameformulation as the at least one compound, or in a separate formulationbefore, during, or after administration of the at least one compound.11. The method of claim 1, wherein two or more compounds selected fromthe group consisting of NCGC00102779-01, NCGC00113159-01,NCGC00104879-01, NCGC00025125-18, NCGC00018238-09, NCGC00112058-01,NCGC00169957-03, NCGC00387651-01, NCGC00179895-03, NCGC00108525-01,NCGC00071621-03, NCGC00131231-01, NCGC00246910-02, NCGC00108581-01,NCGC00107055-01, NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, andNCGC00378623-01, or a prodrug, metabolite, or derivative of any of theforegoing, or a pharmaceutically acceptable salt of any of theforegoing, are administered to the subject, in the same composition orin different compositions.
 12. The method of claim 1, wherein the atleast one compound is administered to the subject in a pharmaceuticalcomposition comprising the at least one compound and a pharmaceuticallyacceptable carrier or diluent.
 13. A method for inhibiting Flavivirusinfection in human or non-human animal cells in vitro or in vivo, saidmethod comprising contacting an effective amount of at least onecompound to a human or non-human animal cell in vitro or in vivo beforeor after exposure of the cell to Flavivirus, wherein the at least onecompound is selected from the group consisting of NCGC00102779-01,NCGC00113159-01, NCGC00104879-01, NCGC00025125-18, NCGC00018238-09,NCGC00112058-01, NCGC00169957-03, NCGC00387651-01, NCGC00179895-03,NCGC00108525-01, NCGC00071621-03, NCGC00131231-01, NCGC00246910-02,NCGC00108581-01, NCGC00107055-01, NCGC00263862-02, NCGC00072088-02,NCGC00357393-02, and NCGC00378623-01, or a prodrug, metabolite, orderivative of any of the foregoing, or a pharmaceutically acceptablesalt of any of the foregoing.
 14. The method of claim 13, wherein theFlavivirus infection is dengue virus infection.
 15. The method of claim13, wherein the cell is contacted with the at least one compound invitro or in vivo after exposure of the cell to the Flavivirus.
 16. Apackaged dosage formulation comprising at least one compound, whereinthe at least one compound is in a pharmaceutically acceptable dosage inone or more packages, packets, or containers, and wherein the at leastone compound is selected from the group consisting of NCGC00102779-01,NCGC00113159-01, NCGC00104879-01, NCGC00025125-18, NCGC00018238-09,NCGC00112058-01, NCGC00169957-03, NCGC00387651-01, NCGC00179895-03,NCGC00108525-01, NCGC00071621-03, NCGC00131231-01, NCGC00246910-02,NCGC00108581-01, NCGC00107055-01, NCGC00263862-02, NCGC00072088-02,NCGC00357393-02, and NCGC00378623-01, or a prodrug, metabolite, orderivative of any of the foregoing, or a pharmaceutically acceptablesalt of any of the foregoing.
 17. The packaged dosage formulation ofclaim 16, comprising two or more of the compounds selected from thegroup consisting of NCGC00102779-01, NCGC00113159-01, NCGC00104879-01,NCGC00025125-18, NCGC00018238-09, NCGC00112058-01, NCGC00169957-03,NCGC00387651-01, NCGC00179895-03, NCGC00108525-01, NCGC00071621-03,NCGC00131231-01, NCGC00246910-02, NCGC00108581-01, NCGC00107055-01,NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01,or a prodrug, metabolite, or derivative of any of the foregoing, or apharmaceutically acceptable salt of any of the foregoing.
 18. A kitcomprising, in one or more containers, at least one compound; andpackaging material, wherein the at least one compound is selected fromthe group consisting of NCGC00102779-01, NCGC00113159-01,NCGC00104879-01, NCGC00025125-18, NCGC00018238-09, NCGC00112058-01,NCGC00169957-03, NCGC00387651-01, NCGC00179895-03, NCGC00108525-01,NCGC00071621-03, NCGC00131231-01, NCGC00246910-02, NCGC00108581-01,NCGC00107055-01, NCGC00263862-02, NCGC00072088-02, NCGC00357393-02, andNCGC00378623-01, or a prodrug, metabolite, or derivative of any of theforegoing, or a pharmaceutically acceptable salt of any of theforegoing.
 19. The kit of claim 18, further comprising instructions forusing the at least one compound or composition for treating orpreventing a Flavivirus infection.
 20. The kit of claim 18, comprisingtwo or more of the compounds selected from the group consisting ofNCGC00102779-01, NCGC00113159-01, NCGC00104879-01, NCGC00025125-18,NCGC00018238-09, NCGC00112058-01, NCGC00169957-03, NCGC00387651-01,NCGC00179895-03, NCGC00108525-01, NCGC00071621-03, NCGC00131231-01,NCGC00246910-02, NCGC00108581-01, NCGC00107055-01, NCGC00263862-02,NCGC00072088-02, NCGC00357393-02, and NCGC00378623-01, or a prodrug,metabolite, or derivative of any of the foregoing, or a pharmaceuticallyacceptable salt of any of the foregoing.